Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients
BackgroundThrombocytopenia is the most notable phenomenon in dengue. Activation status of platelets and interaction of platelets with endothelium contribute towards dengue disease pathogenesis. Platelets are the major cell types known to release extracellular vesicles, especially exosomes in circula...
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1285162/full |
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| author | Sayali Vedpathak Archana Sharma Sonali Palkar Varsha R. Bhatt Vishwanath Chandrashekhar Patil Arjun L. Kakrani AkhileshChandra Mishra Deepak Bhosle Vidya A. Arankalle Shubham Shrivastava |
| author_facet | Sayali Vedpathak Archana Sharma Sonali Palkar Varsha R. Bhatt Vishwanath Chandrashekhar Patil Arjun L. Kakrani AkhileshChandra Mishra Deepak Bhosle Vidya A. Arankalle Shubham Shrivastava |
| author_sort | Sayali Vedpathak |
| collection | DOAJ |
| description | BackgroundThrombocytopenia is the most notable phenomenon in dengue. Activation status of platelets and interaction of platelets with endothelium contribute towards dengue disease pathogenesis. Platelets are the major cell types known to release extracellular vesicles, especially exosomes in circulation. However, the role of platelet derived exosomes (PLT-EXOs) in endothelial dysfunction during dengue infection remains unknown.MethodsIn this study, we recruited 28 healthy subjects and 69 dengue patients categorized as WS- (n=31), WS+ (n=29) and SD (n=9). Platelets were isolated from platelet rich plasma of dengue patients and their activation was assessed by flow cytometry. PLT-EXOs were isolated by ultracentrifugation method. Western blot analyses were performed to characterize the exosomes. Exosome uptake experiment was carried out to see the internalization of exosomes inside endothelial cells (HUVECs). To observe the effect of exosomes on endothelial cells, exosomes were added on HUVECs and expression of adherens and tight junctional proteins were examined by immunofluorescence assay and western blot. Expression levels of vascular injury markers were measured in the culture supernatants of Exosome-HUVEC coculture and sera of dengue patients by MSD-multiplex assay.ResultsAs compared to healthy subjects, CD41/CD61 expression was significantly reduced (p<0.0001) and CD62p expression was significantly increased (p<0.0001) on platelets in dengue patients. PLT-EXOs isolated from the dengue patients showed higher expression of CD63 and CD9 proteins than the healthy subjects. With in-vitro immunofluorescence assays, we illustrated the internalization of PLT-EXOs by the HUVECs and observed disruption of endothelial cell monolayer integrity in the presence of PLT-EXOs from WS+ and SD patients. Furthermore, the significant reduction in the expressions of ZO-2, VE-Cadherin and CD31 in endothelial cells following exposure to PLT-EXOs from the dengue patients provide direct evidence of PLT-EXOs mediated vascular permeability. PLT-EXOs stimulated the release of inflammatory markers CRP, SAA, sVCAM-1 and sICAM-1 in the supernatants of HUVEC cells. Importantly, significantly higher levels of CRP, sVCAM-1 and sICAM-1 in the sera of severe than mild dengue patients (p<0.0001) suggest their role in disease severity.ConclusionsIn summary, our data suggest that PLT-EXOs promote vascular leakage via release of proinflammatory mediators and compromise vascular barrier integrity in dengue patients. |
| first_indexed | 2024-03-08T17:21:39Z |
| format | Article |
| id | doaj.art-49cddce430df4e2ab1f41cd5c8bb7747 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-08T17:21:39Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj.art-49cddce430df4e2ab1f41cd5c8bb77472024-01-03T04:31:44ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-01-011410.3389/fimmu.2023.12851621285162Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patientsSayali Vedpathak0Archana Sharma1Sonali Palkar2Varsha R. Bhatt3Vishwanath Chandrashekhar Patil4Arjun L. Kakrani5AkhileshChandra Mishra6Deepak Bhosle7Vidya A. Arankalle8Shubham Shrivastava9Department of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed to be University), Pune, IndiaDepartment of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed to be University), Pune, IndiaDepartment of Community Medicine, Bharati Vidyapeeth (Deemed to be University) Medical College and Hospital, Pune, IndiaDepartment of Clinical Immunology and Rheumatology, Bharati Vidyapeeth (Deemed to be University) Medical College and Hospital, Pune, IndiaDepartment of Critical Care Medicine, Bharati Vidyapeeth (Deemed to be University) Medical College and Hospital, Pune, IndiaDepartment of Medicine, Dr. D. Y. Patil Medical College Hospital & Research Centre, Dr. D .Y. Patil Vidyapeeth, Pune, IndiaDepartment of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed to be University), Pune, IndiaDepartment of Medicine, Bharati Vidyapeeth (Deemed to be University) Medical College and Hospital, Pune, IndiaDepartment of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed to be University), Pune, IndiaDepartment of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed to be University), Pune, IndiaBackgroundThrombocytopenia is the most notable phenomenon in dengue. Activation status of platelets and interaction of platelets with endothelium contribute towards dengue disease pathogenesis. Platelets are the major cell types known to release extracellular vesicles, especially exosomes in circulation. However, the role of platelet derived exosomes (PLT-EXOs) in endothelial dysfunction during dengue infection remains unknown.MethodsIn this study, we recruited 28 healthy subjects and 69 dengue patients categorized as WS- (n=31), WS+ (n=29) and SD (n=9). Platelets were isolated from platelet rich plasma of dengue patients and their activation was assessed by flow cytometry. PLT-EXOs were isolated by ultracentrifugation method. Western blot analyses were performed to characterize the exosomes. Exosome uptake experiment was carried out to see the internalization of exosomes inside endothelial cells (HUVECs). To observe the effect of exosomes on endothelial cells, exosomes were added on HUVECs and expression of adherens and tight junctional proteins were examined by immunofluorescence assay and western blot. Expression levels of vascular injury markers were measured in the culture supernatants of Exosome-HUVEC coculture and sera of dengue patients by MSD-multiplex assay.ResultsAs compared to healthy subjects, CD41/CD61 expression was significantly reduced (p<0.0001) and CD62p expression was significantly increased (p<0.0001) on platelets in dengue patients. PLT-EXOs isolated from the dengue patients showed higher expression of CD63 and CD9 proteins than the healthy subjects. With in-vitro immunofluorescence assays, we illustrated the internalization of PLT-EXOs by the HUVECs and observed disruption of endothelial cell monolayer integrity in the presence of PLT-EXOs from WS+ and SD patients. Furthermore, the significant reduction in the expressions of ZO-2, VE-Cadherin and CD31 in endothelial cells following exposure to PLT-EXOs from the dengue patients provide direct evidence of PLT-EXOs mediated vascular permeability. PLT-EXOs stimulated the release of inflammatory markers CRP, SAA, sVCAM-1 and sICAM-1 in the supernatants of HUVEC cells. Importantly, significantly higher levels of CRP, sVCAM-1 and sICAM-1 in the sera of severe than mild dengue patients (p<0.0001) suggest their role in disease severity.ConclusionsIn summary, our data suggest that PLT-EXOs promote vascular leakage via release of proinflammatory mediators and compromise vascular barrier integrity in dengue patients.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1285162/fulldengue virusplateletsexosomesplatelet-derived exosomesvascular inflammationsVCAM-1 |
| spellingShingle | Sayali Vedpathak Archana Sharma Sonali Palkar Varsha R. Bhatt Vishwanath Chandrashekhar Patil Arjun L. Kakrani AkhileshChandra Mishra Deepak Bhosle Vidya A. Arankalle Shubham Shrivastava Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients Frontiers in Immunology dengue virus platelets exosomes platelet-derived exosomes vascular inflammation sVCAM-1 |
| title | Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients |
| title_full | Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients |
| title_fullStr | Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients |
| title_full_unstemmed | Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients |
| title_short | Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients |
| title_sort | platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients |
| topic | dengue virus platelets exosomes platelet-derived exosomes vascular inflammation sVCAM-1 |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1285162/full |
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